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Chaperone proteins as single component reagents to assess antibody nonspecificity

Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are...

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Autores principales: Kelly, Ryan L., Geoghegan, James C., Feldman, Jared, Jain, Tushar, Kauke, Monique, Le, Doris, Zhao, Jessie, Wittrup, K. Dane
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Taylor & Francis 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627595/
https://www.ncbi.nlm.nih.gov/pubmed/28745541
http://dx.doi.org/10.1080/19420862.2017.1356529
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author Kelly, Ryan L.
Geoghegan, James C.
Feldman, Jared
Jain, Tushar
Kauke, Monique
Le, Doris
Zhao, Jessie
Wittrup, K. Dane
author_facet Kelly, Ryan L.
Geoghegan, James C.
Feldman, Jared
Jain, Tushar
Kauke, Monique
Le, Doris
Zhao, Jessie
Wittrup, K. Dane
author_sort Kelly, Ryan L.
collection PubMed
description Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are often ill-defined, and can suffer from issues such as lot-to-lot variability. In this study, we discovered in immunoprecipitation experiments that certain chaperones are present in one of these mixtures;we then explored the use of recombinant chaperone proteins as well-characterized agents to predict antibody nonspecificity. Antibody binding to the heat shock proteins HSP70, HSP90, or trigger factor all served as predictors of cross-interaction propensity, with HSP90 providing the greatest ability to predict antibody clearance rates in mouse. Individual chaperone binding correlates surprisingly closely with binding to complex cell extracts, with the exception of a few “false negatives” (assuming a complex cell extract as the “true” value). As defined reagents, these chaperone reagents present advantages for high throughput assays of nonspecificity.
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spelling pubmed-56275952017-10-12 Chaperone proteins as single component reagents to assess antibody nonspecificity Kelly, Ryan L. Geoghegan, James C. Feldman, Jared Jain, Tushar Kauke, Monique Le, Doris Zhao, Jessie Wittrup, K. Dane MAbs Short Communication Early stage assays that evaluate monoclonal antibody drug-like properties serve as valuable tools for selection of lead candidates. One liability for clinical development, off-target reactivity, is often assessed by binding to a mixture or panel of noncognate proteins. While robust, these mixes are often ill-defined, and can suffer from issues such as lot-to-lot variability. In this study, we discovered in immunoprecipitation experiments that certain chaperones are present in one of these mixtures;we then explored the use of recombinant chaperone proteins as well-characterized agents to predict antibody nonspecificity. Antibody binding to the heat shock proteins HSP70, HSP90, or trigger factor all served as predictors of cross-interaction propensity, with HSP90 providing the greatest ability to predict antibody clearance rates in mouse. Individual chaperone binding correlates surprisingly closely with binding to complex cell extracts, with the exception of a few “false negatives” (assuming a complex cell extract as the “true” value). As defined reagents, these chaperone reagents present advantages for high throughput assays of nonspecificity. Taylor & Francis 2017-07-26 /pmc/articles/PMC5627595/ /pubmed/28745541 http://dx.doi.org/10.1080/19420862.2017.1356529 Text en © 2017 The Author(s). Published with license by Taylor & Francis Group, LLC http://creativecommons.org/licenses/by-nc-nd/4.0/ This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivatives License (http://creativecommons.org/licenses/by-nc-nd/4.0/), which permits non-commercial re-use, distribution, and reproduction in any medium, provided the original work is properly cited, and is not altered, transformed, or built upon in any way.
spellingShingle Short Communication
Kelly, Ryan L.
Geoghegan, James C.
Feldman, Jared
Jain, Tushar
Kauke, Monique
Le, Doris
Zhao, Jessie
Wittrup, K. Dane
Chaperone proteins as single component reagents to assess antibody nonspecificity
title Chaperone proteins as single component reagents to assess antibody nonspecificity
title_full Chaperone proteins as single component reagents to assess antibody nonspecificity
title_fullStr Chaperone proteins as single component reagents to assess antibody nonspecificity
title_full_unstemmed Chaperone proteins as single component reagents to assess antibody nonspecificity
title_short Chaperone proteins as single component reagents to assess antibody nonspecificity
title_sort chaperone proteins as single component reagents to assess antibody nonspecificity
topic Short Communication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5627595/
https://www.ncbi.nlm.nih.gov/pubmed/28745541
http://dx.doi.org/10.1080/19420862.2017.1356529
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